ABSTRACT
We have cloned and sequenced the cDNA encoding an immunoreactive protein from the pathogenic fungus Coccidioides immitis which stimulates human T cells and has been associated with protective vaccines in mice. The transcript contained an open reading frame encoding 194 amino acids with a calculated molecular weight of 19.5 kDa, a 151 base 5' untranslated region (UTR), and a 468 base 3'UTR. A four member repeat motif, usually thr-ala-glu-pro, exists for amino acids 98 through 141. Deduced amino acid sequence derived from the cDNA was identical with previously determined internal amino acid sequence from the native protein, and goat antiserum raised against the purified fungal protein reacted with an inducible fusion protein translated from this cDNA. Using this cDNA to produce recombinant protein will allow direct testing of its role in human immunity to coccidioidomycosis and may lead to new diagnostic tests.
Subject(s)
Antigens, Fungal/genetics , Coccidioides/genetics , Fungal Proteins/genetics , Amino Acid Sequence , Base Sequence , DNA Primers/chemistry , DNA, Complementary/genetics , Fungal Proteins/immunology , Genes, Fungal , Humans , Molecular Sequence Data , Peptides/genetics , Peptides/immunology , Proline-Rich Protein Domains , RNA, Fungal/geneticsABSTRACT
We performed antifungal susceptibility tests with cilofungin (LY121019), amphotericin B, and flucytosine against 38 strains of yeasts from patients with esophagitis or fungemia either before, during, or after treatment with cilofungin. Tests were performed using a macrobroth dilution method similar to that proposed by the National Committee for Clinical Laboratory Standards (M27-P) and two microbroth methods. For cilofungin and amphotericin B, minimum inhibitory concentrations from microbroth tests using Antibiotic Medium 3 (AM3) were systematically lower than results from the other two methods that utilized RPMI-1640 medium (RPMI). AM3 did not provide any greater degree of in vitro correlation with clinical results than did RPMI. We conclude that cilofungin and possibly other congeners of the echinocandin class of antifungal agents can effectively be studied using the proposed National Committee for Clinical Laboratory Standards method.
